The present invention relates to agents that act to antagonize the action of the glucagon peptide hormone. It relates particularly to non-peptide glucagon antagonists or inverse agonists.
Glucagon is a key hormonal agent that, in cooperation with insulin, mediates homeostatic regulation of the amount of glucose in the blood. Glucagon primarily acts by stimulating certain cells (mostly liver cells) to release glucose when blood glucose levels fall. The action of glucagon is opposed by insulin which stimulates cells to take up and store glucose whenever blood glucose levels rise. Both glucagon and insulin are peptide hormones.
Glucagon is produced in the alpha islet cells and insulin in the beta islet cells of the pancreas. Diabetes mellitus, the common disorder of glucose metabolism, is characterized by hyperglycemia, and can present as type I, insulin-dependent, or type II, a form that is non-insulin-dependent in character. Subjects with type I diabetes are hyperglycemic and hypoinsulinemic, and the conventional treatment for this form of the disease is to provide insulin. However, in some patients with type I or II diabetes, absolute or relative elevated glucagon levels have been shown to contribute to the hyperglycemic state. Both in healthy animals as well as in animal models of type I and II, removal of circulating glucagon with selective and specific anti-bodies has resulted in reduction of the glycemic level (Brand et al. Diabetologia 37, 985 (1994); Diabetes 43, [suppl 1], 172A (1994); Am J Physiol 269, E469-E477 (1995); Diabetes 44 [suppl 1], 134A (1995); Diabetes 45, 1076 (1996)). These studies suggest that glucagon suppression or an action antagonistic to glucagon could be a useful adjunct to conventional antihyperglycemia treatment of diabetes. The action of glucagon can be suppressed by providing an antagonist or an inverse agonist, substances that inhibit or prevent glucagon induced response. The antagonist can be peptide or non-peptide in nature. Native glucagon is a 29 amino acid-containing peptide having the sequence:
His-Ser-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Ser-Arg-Arg-Ala-Gln-Asp-Phe-Val-Gln-Trp-Leu-Met-Asn-Thr-NH2.
Glucagon exerts its action by binding to and activating its receptor, which is part of the Glucagon-Secretin branch of the 7-transmembrane G-protein coupled receptor family (Jelinek et al. Science 259, 1614, (1993)). The receptor functions by activation of the adenylyl cyclase second messenger system and the result is an increase in cAMP levels.
Several publications disclose peptide antagonists. Probably, the most thoroughly characterized antagonist is DesHis1[Glu9]-glucagon amide (Unson et al., Peptides 10, 1171 (1989); Post et al., Proc. Natl. Acad. Sci. USA 90, 1662 (1993)). Other antagonists are eg DesHis1,Phe6[Glu9]-glucagon amide (Azizh et al., Bioorganic and Medicinal Chem. Lett. 16, 1849 (1995)) or NLeu9,Ala11,16-glucagon amide (Unson et al., J. Biol. Chem. 269(17), 12548 (1994)).
Peptide antagonists of peptide hormones are often quite potent; however, they are defective as drugs because of degradation by physiological enzymes, and poor biodistribution. Therefore, non-peptide antagonists of the peptide hormones are preferred. Among the non-peptide glucagon antagonists, a quinoxaline derivative, (2-styryl-3-[3-(dimethylamino)propylmethyl-amino]-6,7-dichloroquinoxaline was found to displace glucagon from the rat liver receptor (Collins, J. L. et al. (1992) Bioorganic and Medicinal Chemistry Letters 2(9):915-918). West, R. R. et al. (1994), WO 94/14426 discloses use of skyrin, a natural product comprising a pair of linked 9,10-anthracenedione groups, and its synthetic analogues, as glucagon antagonists. Anderson, P. L., U.S. Pat. No. 4,359,474 discloses the glucagon antagonistic properties of 1-phenyl pyrazole derivatives. Barcza, S., U.S. Pat. No. 4,374,130, discloses substituted disilacyclohexanes as glucagon antagonists. WO 98/04528 (Bayer Corporation) discloses substituted pyridines and biphenyls as glucagon antagonists. Furthermore, WO 97/16442 (Merck and Co., Inc.) discloses substituted pyridyl pyrroles as glucagon antagonists and WO 98/21957 (Merck and Co., Inc.) discloses 2,4-diaryl-5-pyridylimidazoles as glucagon antagonists. These glucagon antagonists differ structurally from the present compounds.
The following is a detailed definition of the terms used to describe the compounds of the invention:
xe2x80x9cHalogenxe2x80x9d designates an atom selected from the group consisting of F, Cl, Br or I.
The term xe2x80x9calkylxe2x80x9d in the present context designates a hydrocarbon chain or a ring that is either saturated or unsaturated (containing one or more double or triple bonds where feasible) of from 1 to 10 carbon atoms in either a linear or branched or cyclic configuration. Thus, alkyl includes for example n-octyl, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, allyl, propargyl, 2-hexynyl, cyclopropyl, cyclopropylmethyl, cyclopentyl, cyclohexyl, cyclooctyl, 4-cyclohexylbutyl, and the like.
Further non-limiting examples are sec-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl, n-heptyl, n-nonyl, n-decyl, vinyl, 1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 1-hexenyl, 3-hexenyl, 2,4-hexadienyl, 5-hexenyl, 1-heptenyl, 2,4-heptadienyl, 1-octenyl, 2,4-octadienyl, ethynyl, 1-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 3-hexynyl, 2,4-hexadiynyl, 5-hexynyl, 1-hepynyl, 1-octynyl, 2-decynyl, cyclobutyl, cyclopentyl, 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cyclohexenyl, 2-cyclohexenyl, 3-cyclohexenyl, 2-cyclopropylethyl, cyclobutylmethyl, 2-cyclobutylethyl, cyclohexenylmethyl, 4-cyclohexyl-2-butenyl, 4-(1-cyclohexenyl)-vinyl and the like.
The term xe2x80x9clower alkylxe2x80x9d designates a hydrocarbon moiety specified above, of from 1 to 6 carbon atoms.
xe2x80x9cArylxe2x80x9d means an aromatic ring moiety, for example: phenyl, naphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyrimidinyl, pyrazolyl, imidazolyl, pyrazinyl, pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, thiazolyl, isothiazolyl, tetrazolyl, 1-H-tetrazol-5-yl, indolyl, quinolyl, quinazolinyl, benzofuryl, benzothiophenyl (thianaphthenyl) and the like.
Further non-limiting examples are biphenyl, anthracenyl, phenanthrenyl, fluorenyl, indenyl, 1,2,3,4-tetrahydronaphthyl, 2,3-dihydrobenzofuryl, triazolyl, pyranyl, thiadiazinyl, isoindolyl, indazolyl, 1,2,5-oxadiazolyl, 1,2,5-thiadiazolyl, benzothienyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, purinyl, quinolizinyl, isoquinolyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, pyrrolinyl, pyrazolinyl, indolinyl, pyrrolidinyl, piperidinyl and the like.
The aryl moieties are optionally substituted by one or more substituents, for example selected from the group consisting of F, Cl, I, and Br; lower alkyl; lower alkanoyl such as formyl, acetyl, propionyl, butyryl, valeryl, hexanoyl and the like; xe2x80x94OH; xe2x80x94NO2; xe2x80x94CN; xe2x80x94CO2H; xe2x80x94O-lower alkyl; aryl; aryl-lower alkyl; xe2x80x94CO2CH3; xe2x80x94CONH2; xe2x80x94OCH2CONH2; xe2x80x94NH2; xe2x80x94N(CH3)2; xe2x80x94SO2NH2; xe2x80x94OCHF2; xe2x80x94CF3; xe2x80x94OCF3 and the like. A further non-limiting example is xe2x80x94NHxe2x80x94(Cxe2x95x90S)xe2x80x94NH2.
Such aryl moieties may also be substituted by two substituents forming a bridge, for example xe2x80x94OCH2Oxe2x80x94.
xe2x80x9cAryl-lower alkylxe2x80x9d means a lower alkyl as defined above, substituted by an aryl, for example: 
The aryl group is optionally substituted as described above.
The present invention is based on the unexpected observation that compounds having a selected nitrogen-bearing central motif and the general structural features disclosed below antagonize the action of glucagon.
Accordingly, the invention is concerned with compounds of the general formula I: 
wherein:
R1 and R2 independently are hydrogen or lower alkyl or together form a valence bond;
R3 and R4independently are hydrogen or lower alkyl;
n is 0, 1, 2 or 3;
m is 0 or 1;
X is  greater than Cxe2x95x90O,  greater than Cxe2x95x90S,  greater than Cxe2x95x90NR5 or  greater than SO2;
wherein R5 is hydrogen, lower alkyl, aryl-lower alkyl or xe2x80x94OR6;
wherein R6 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
A is 
xe2x80x83wherein:
R7 is hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR11, xe2x80x94NR11R12, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SO2NR11R12, xe2x80x94SR11, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OSO2R11, xe2x80x94CONR11R12, xe2x80x94OCH2CONR11R12, xe2x80x94CH2OR11, xe2x80x94CH2NR11R12, xe2x80x94OCOR11, xe2x80x94CO2R13 or xe2x80x94OSO2CF3;
R8 and R9 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR11, xe2x80x94NR11R12, lower alkyl, aryl, xe2x80x94SCF3, xe2x80x94SR11, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OSO2R11, xe2x80x94CONR11R12, xe2x80x94CH2OR11, xe2x80x94CH2NR11R12, xe2x80x94OCOR11, xe2x80x94CO2R13 or xe2x80x94OSO2CF3, or R8 and R9 together form a bridge xe2x80x94OCH2Oxe2x80x94 or xe2x80x94OCH2CH2Oxe2x80x94;
wherein R11 and R12 independently are hydrogen, xe2x80x94COR13, xe2x80x94SO2R13, lower alkyl or aryl;
wherein R13 is hydrogen, lower alkyl, aryl-lower alkyl or aryl; and
R10 is hydrogen, lower alkyl, aryl-lower alkyl or aryl; B is 
or a valence bond;
xe2x80x83wherein:
R14 and R15 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94O(CH2)lCF3, xe2x80x94NO2, xe2x80x94OR16, xe2x80x94NR16R17, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR16, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR16R17, xe2x80x94(CH2)lCONR16R17, xe2x80x94O(CH2)lCONR16R17, xe2x80x94(CH2)lCOR16, xe2x80x94(CH2)lCOR16, xe2x80x94(CH2)lOR16, xe2x80x94O(CH2)lOR16, xe2x80x94(CH2)lNR16R17, xe2x80x94O(CH2)lNR16R17, xe2x80x94OCOR16, xe2x80x94CO2R18, xe2x80x94O(CH2)lCO2R18, xe2x80x94O(CH2)lCN, xe2x80x94O(CH2)lCl , or R14 and R15 together form a bridge xe2x80x94O(CH)lOxe2x80x94 or xe2x80x94(CH2)lxe2x80x94;
wherein l is 1, 2, 3 or 4;
R16 and R17 independently are hydrogen, xe2x80x94COR18, xe2x80x94SO2R18, lower alkyl, aryl, or R16 and R17 together form a cyclic alkyl bridge containing from 2 to 7 carbon atoms;
wherein R18 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
W is xe2x80x94Nxe2x95x90 or xe2x80x94CR19xe2x95x90;
Y is xe2x80x94Nxe2x95x90 or xe2x80x94CR20xe2x95x90;
Z is xe2x80x94Nxe2x95x90 or xe2x80x94CR21xe2x95x90;
V is xe2x80x94Nxe2x95x90 or xe2x80x94CR22xe2x95x90; and
Q is xe2x80x94NR23xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
wherein:
R19, R20, R21 and R22 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR24, xe2x80x94NR24R25, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR24, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR24R25, xe2x80x94CH2CONR24R25, xe2x80x94OCH2CONR24R25, xe2x80x94CH2OR24, xe2x80x94CH2NR24R25, xe2x80x94OCOR24 or xe2x80x94CO2R24, or R19 and R20, R20 and R21, or R21, and R22 together form a bridge xe2x80x94OCH2Oxe2x80x94;
wherein R24 and R25 independently are hydrogen, xe2x80x94COR26, xe2x80x94SO2R26, lower alkyl, aryl or aryl-lower alkyl;
wherein R26 is hydrogen, lower alkyl, aryl or aryl-lower alkyl; and
R23 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
K is 
xe2x80x83wherein:
R3a, R3b, R4a and R4b independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94CCH2CF3, xe2x80x94NO2, xe2x80x94OR24, xe2x80x94NR24aR25a, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR24a, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR24aR25a, xe2x80x94CH2CONR24aR25a, xe2x80x94OCH2CONR24aR25a, xe2x80x94CH2OR24a, xe2x80x94CH2NR24aR25a, xe2x80x94OCOR24a or xe2x80x94CO2R24a;
wherein R24a and R25a independently are hydrogen, xe2x80x94COR26a, xe2x80x94SO2R26a, lower alkyl, aryl or aryl-lower alkyl;
wherein R26a is hydrogen, lower alkyl, aryl or aryl-lower alkyl; or
R3a and R3b, R4a and R4b, or R3a and R4b together form a bridge xe2x80x94(CH2)ixe2x80x94;
wherein i is 1, 2, 3 or 4;
a, b, c and d independently are 0, 1, 2, 3 or 4;
e, f and p independently are 0 or 1;
q is 0, 1 or 2; and
L and M independently are xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94CHxe2x95x90CHxe2x80x94, xe2x80x94Cxe2x89xa1Cxe2x80x94, xe2x80x94NR5axe2x80x94, xe2x80x94CH2NR5axe2x80x94, xe2x80x94COxe2x80x94, xe2x80x94OCOxe2x80x94, xe2x80x94COOxe2x80x94, xe2x80x94CONR5axe2x80x94, CONR5bxe2x80x94, xe2x80x94NR5aCOxe2x80x94, xe2x80x94SOxe2x80x94, xe2x80x94SO2xe2x80x94, xe2x80x94OSO2xe2x80x94, xe2x80x94SO2NR5axe2x80x94, xe2x80x94NR5aSO2xe2x80x94, xe2x80x94NR5aCONR5bxe2x80x94, xe2x80x94CONR5aNR5bxe2x80x94, xe2x80x94NR5aCSNR5bxe2x80x94, xe2x80x94OCONR5bxe2x80x94, xe2x80x94CH2CONR5bxe2x80x94, xe2x80x94OCH2CONR5bxe2x80x94,
xe2x80x94P(O)(OR5a)Oxe2x80x94, xe2x80x94NR5aC(O)Oxe2x80x94 or 
xe2x80x83wherein R5a and R5b independently are hydrogen, lower alkyl, xe2x80x94OH, xe2x80x94(CH2)kxe2x80x94OR6axe2x80x94, xe2x80x94COR6a, xe2x80x94(CH2)kxe2x80x94CH(OR6a)2, xe2x80x94(CH2)kxe2x80x94CN, xe2x80x94(CH2)kxe2x80x94NR6aR6bxe2x80x94, aryl, aryl-lower alkyl, xe2x80x94(CH2)gxe2x80x94COOR43 or xe2x80x94(CH2)gxe2x80x94CF3;
wherein k is 1, 2, 3 or 4;
R6a and R6bindependently are hydrogen, lower alkyl, aryl or aryl-lower alkyl;
g is 0, 1, 2, 3 or 4;
R43 is hydrogen or lower alkyl;
Gxe2x80x3 is xe2x80x94OCH2COxe2x80x94, xe2x80x94CH2COxe2x80x94, xe2x80x94COxe2x80x94 or a valence bond; and
Exe2x80x3 is xe2x80x94CH2xe2x80x94, xe2x80x94CH2CH2xe2x80x94, xe2x80x94CHxe2x95x90CHxe2x80x94, xe2x80x94CH2NHxe2x80x94 or xe2x80x94CH2CH2NHxe2x80x94;
D is hydrogen, 
wherein:
r is 0 or 1 ;
s is 0, 1, 2 or 3;
E, Exe2x80x2, F, G and Gxe2x80x3 independently are xe2x80x94CHR38xe2x80x94,  greater than Cxe2x95x90O,  greater than NR39, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
Fxe2x80x2 is  greater than CR38xe2x80x94 or  greater than Nxe2x80x94;
Yxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR32xe2x95x90;
Zxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR33xe2x95x90;
Vxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR34xe2x95x90;
Wxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR35xe2x95x90; and
Qxe2x80x2 is xe2x80x94NR36xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
wherein:
R27, R28, R32, R33, R34 and R35 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94O(CH2)yCF3, xe2x80x94(CH2)yNHCOCF3, xe2x80x94NO2, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR29, xe2x80x94CHF2, xe2x80x94OH2, xe2x80x94OCF2CHF2, xe2x80x94OSO2R29, xe2x80x94OSO2CF3, xe2x80x94(CH2)yCONR29R30, xe2x80x94O(CH2)yCONR29R30, xe2x80x94(CH2)yOR29, xe2x80x94(CH2)yNR29R30, xe2x80x94OCOR29, xe2x80x94COR29 or xe2x80x94CO2R29; or
R27 and R28, R32 and R33, R34 and R34 or R34 and R35 together form a bridge xe2x80x94OCH2)yOxe2x80x94,
wherein y is 0, 1, 2, 3 or 4; and
R29 and R30 independently are hydrogen, xe2x80x94CNR31, xe2x80x94CO2R31, xe2x80x94SO2R31, lower alkyl, aryl or aryl-lower alkyl;
wherein R31 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
R36 and R39 independently are hydrogen, lower alkyl, aryl or aryl-lower alkyl; and
R38 is hydrogen, xe2x80x94OR40, xe2x80x94NR40R41, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR40, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94CONR40R41, xe2x80x94(CH2)xCONR40OR41, xe2x80x94O(CH2)xCONR40R41, xe2x80x94(CH2)xOR40, xe2x80x94(CH2)xNR40OR41, xe2x80x94OCOR40 or xe2x80x94CO2R40;
wherein x is 1, 2, 3 or 4;
R40 and R41 independently are hydrogen, xe2x80x94COR42, xe2x80x94SO2R42, lower alkyl, aryl or aryl-lower alkyl;
wherein R42 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
as well as any optical or geometric isomer or tautomeric form thereof including mixtures of these or a pharmaceutically acceptable salt thereof.
Where the formulae for B make it possible, R19, R20, R21, R22 and R23 may alternatively be replaced by R14 or R15, respectively. In such case eg W may be selected from xe2x80x94Nxe2x95x90, xe2x80x94CR19xe2x80x94 and xe2x80x94C14xe2x80x94.
Similarly, where the formulae for D make it possible, R32, R33, R34, R35, R36, R38 and R39 may alternatively be replaced by R27 or R28, respectively. In such case eg E may be selected from xe2x80x94CHR38xe2x80x94,  greater than Cxe2x95x90O,  greater than NR39, xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94CHR27xe2x80x94 and  greater than NR27.
In a preferred embodiment the invention relates to compounds of the following general formula II. 
wherein A, B, K, D, R3, R4, n and m are as defined for formula I.
In another preferred embodiment the invention relates to compounds of the following general formula III: 
wherein A, B, K, D, R3, R4, n and m are as defined for formula I.
In still another preferred embodiment the invention relates to compounds of the following formula IV: 
wherein A , B, K, D, R3, R4, n and m are as defined for formula I.
In the compounds of the above formulae I to IV the following substituents are preferred:
R3 is preferably hydrogen.
R4 is preferably hydrogen.
A is preferably selected from the group consisting of: 
xe2x80x83wherein R7, R8, R9 and R10 are as defined for formula I.
A is more preferably 
xe2x80x83wherein R7, R8 and R9 are as defined for formula I.
In the above embodiments of A, R7 is preferably halogen, lower alkyl, xe2x80x94OH, xe2x80x94NO2, xe2x80x94CN, xe2x80x94CO2H, xe2x80x94O-lower alkyl, aryl, aryl-lower alkyl, xe2x80x94CO2CH3, xe2x80x94CONH2, xe2x80x94OCH2CONH2, xe2x80x94NH2, xe2x80x94N(CH3)2, xe2x80x94SO2NH2, xe2x80x94OCHF2, xe2x80x94CF3 or xe2x80x94OCF3.
Preferably, R8 and R9 are independently hydrogen, halogen, xe2x80x94OH, xe2x80x94NO2, xe2x80x94NH2, xe2x80x94CN, xe2x80x94OCF3, xe2x80x94SCF3, xe2x80x94CF3, xe2x80x94OCH2CF3, xe2x80x94O-lower alkyl such as methoxy and ethoxy, lower alkyl such as methyl and ethyl, or phenyl, and R10 is hydrogen, lower alkyl or phenyl.
More preferably, R8 and R9 are independently selected from hydrogen, halogen such as xe2x80x94F and xe2x80x94Cl, xe2x80x94O-lower alkyl such as methoxy and ethoxy, xe2x80x94NH2, xe2x80x94CN or xe2x80x94NO2, and R10 is hydrogen.
In a particularly preferred embodiment A is 
wherein R8 and R9 independently are hydrogen, halogen, xe2x80x94OH, xe2x80x94NO2, xe2x80x94NH2, xe2x80x94CN, xe2x80x94OCF3, xe2x80x94SCF3, xe2x80x94CF3, xe2x80x94OCH2CF3, xe2x80x94O-lower alkyl such as methoxy and ethoxy, lower alkyl such as methyl and ethyl, or phenyl, preferably hydrogen, halogen such as xe2x80x94F and xe2x80x94Cl, xe2x80x94O-lower alkyl such as methoxy and ethoxy, xe2x80x94NH2, xe2x80x94CN or xe2x80x94NO2.
In a further particularly preferred embodiment A is 
wherein R8 is hydrogen, halogen such as xe2x80x94F or xe2x80x94Cl, xe2x80x94O-lower alkyl such as xe2x80x94OCH3 or xe2x80x94OC2H5, xe2x80x94NH2, xe2x80x94CN or xe2x80x94NO2; and R9 is hydrogen or halogen such as xe2x80x94F or xe2x80x94Cl.
In a preferred embodiment R8 is halogen and R9 is hydrogen.
In still a preferred embodiment the invention relates to compounds of the following formula V: 
wherein R4, B, K, D and m are as defined for formula I and R8 and R9 are as defined for formula I and preferably as defined for the preferred embodiments of A above.
B is preferably: 
wherein V, W, Z, Y and Q are as defined for formula I; and
R14 and R15 independently are hydrogen, halogen, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OR16, xe2x80x94NR16R17, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94OSO2CF3, xe2x80x94CONR16R17, xe2x80x94CH2OR16, xe2x80x94CH2NR16R17, xe2x80x94OCOR16 or xe2x80x94CO2R18; or R14 and R15 together form a bridge xe2x80x94OCH2Oxe2x80x94 or xe2x80x94(CH2)lxe2x80x94;
wherein l, R16, R17 and R18 are as defined for formula I.
Q is preferably xe2x80x94Oxe2x80x94 or xe2x80x94NHxe2x80x94.
Particularly preferred compounds are those in which B is 
wherein V, W, Z, Y and Q are as defined for formula I; and
R14 and R15 independently are hydrogen, halogen, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OR16, xe2x80x94NR16R17, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94OSO2CF3, xe2x80x94CONR16R17, xe2x80x94CH2OR16, xe2x80x94CH2NR16R17, xe2x80x94OCOR16 or xe2x80x94CO2R18; or R14 and R15 together form a bridge xe2x80x94OCH2Oxe2x80x94 or xe2x80x94(CH2)lxe2x80x94;
wherein l, R16, R17 and R18 are as defined for formula I.
Still more preferred are compounds of the following formula VI: 
as well as compounds of the following formula VII: 
as well as compounds of the general formulae VIIIa or VIIIb: 
wherein R14 and R15 independently are hydrogen, halogen, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OR16, xe2x80x94NR16R17, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94OSO2CF3, xe2x80x94CONR16R17, xe2x80x94CH2OR16, xe2x80x94CH2NR16R17, xe2x80x94OCOR16 or xe2x80x94CO2R18; or R14 and R15 together form a bridge xe2x80x94OCH2Oxe2x80x94 or xe2x80x94(CH2)lxe2x80x94;
wherein l, R16, R14 and R18 are as defined for formula I;
K, D and m are as defined for formula I; and
R8 and R9 are as defined for formula I and preferably as defined for the preferred embodiments of A above.
In the above formulae VI, VII and VIII, R14 and R15 are preferably independently hydrogen, halogen, lower alkyl, aryl such as phenyl, or xe2x80x94O-lower alkyl such as methoxy.
In the above formulae VI and VII, K is preferably bound in para-position and in the above formulae VIIIa and VIIIb, K is preferably bound at the nitrogen atom of the indole group.
K is preferably selected from the group consisting of: 
wherein R3a, R3b, R4a, R4b, R5a, R5b, a, b, c, d, p and q are as defined for formula I.
More preferably, K is selected from the group consisting of: 
wherein R3a, R3b, R4a, R4b, R5a, R5b, a, b, c, d, p and q are as defined for formula I.
In a further preferred embodiment K is selected from the group consisting of: 
wherein R3a, R3b, R4a, R4b, R5a, R5b, b, c, d, p and q are as defined for formula I.
In the above embodiments of K, R5a and R5b are preferably independently hydrogen, lower alkyl, xe2x80x94OH, xe2x80x94(CH2)kOR6a, aryl, aryl-lower alkyl, xe2x80x94CH2CF3, xe2x80x94(CH2)gCOOR43, xe2x80x94COOR43, xe2x80x94(CH2)kxe2x80x94CN or xe2x80x94(CH2)kNR6aR6b wherein g, k, R43, R6a and R5b are as defined for formula I.
Preferably, g and k are independently 1, 2 or 3,and R6a and R6b are independently hydrogen, lower alkyl such as methyl or ethyl, or aryl such as phenyl,
In the above embodiments of K, R3a and R3b are preferably independently hydrogen, halogen, xe2x80x94OH, xe2x80x94O-lower alkyl, xe2x80x94COO-lower alkyl, lower alkyl or aryl-lower alkyl.
In the above embodiments of K, R4a and R4b are preferably independently hydrogen, xe2x80x94CN, xe2x80x94CONH2, xe2x80x94(CH2)xe2x80x94N(CH3)2, xe2x80x94O-lower alkyl, xe2x80x94CH2OH, xe2x80x94CH2O-aryl, xe2x80x94N(CH3)2, xe2x80x94OH, xe2x80x94CO2-lower alkyl or lower alkyl.
D is preferably hydrogen, 
wherein s, r, R27, R28, Vxe2x80x2, Yxe2x80x2, Qxe2x80x2, Zxe2x80x2, Wxe2x80x2, E, Exe2x80x2, F, Fxe2x80x2, G and Gxe2x80x2 are as defined for formula I.
In still a further preferred embodiment D is hydrogen, 
wherein s, r, R27, R28, Vxe2x80x2, Yxe2x80x2, Qxe2x80x2, Zxe2x80x2, Wxe2x80x2, E, Exe2x80x2, F, Fxe2x80x2, G and Gxe2x80x2 are as defined for formula I.
D is more preferably hydrogen, 
wherein E and Exe2x80x2 independently are  greater than CHR38,  greater than NR39 or xe2x80x94Oxe2x80x94; F, G and Gxe2x80x2 independently are  greater than CHR38,  greater than Cxe2x95x90O or  greater than NR39; Fxe2x80x2 is  greater than CR38xe2x80x94 or  greater than Nxe2x80x94; and s, r, R27, R28, R38, R39, Vxe2x80x2, Yxe2x80x2, Zxe2x80x2, Qxe2x80x2 and Wxe2x80x2 are as defined for formula I.
R27 and R28 are preferably independently hydrogen; halogen such as xe2x80x94Cl, xe2x80x94Br or xe2x80x94F; xe2x80x94CF3; xe2x80x94OCF3; xe2x80x94OCHF2; xe2x80x94OCH2CF3; xe2x80x94(CH2)yNHCOCF3; xe2x80x94NHCOCF3; xe2x80x94CN; xe2x80x94NO2; xe2x80x94COR29, xe2x80x94COOR29, xe2x80x94(CH2)yOR29 or xe2x80x94OR29 wherein R29 is hydrogen, aryl or lower alkyl and y is 1, 2, 3 or 4; lower alkyl such as methyl, ethyl, 2-propenyl, isopropyl, tert-butyl or cyclohexyl; lower alkylthio; xe2x80x94SCF3; aryl such as phenyl; xe2x80x94(CH2)yNR29R30 or xe2x80x94NR29R30 wherein R29 and R30 independently are hydrogen, xe2x80x94COO-lower alkyl or lower alkyl and y is 1, 2, 3 or 4; or xe2x80x94CONH2; or R27 and R28 together form a bridge xe2x80x94OCH2Oxe2x80x94; R38 is hydrogen; xe2x80x94OCHF2; xe2x80x94OR40 wherein R40 is hydrogen or lower alkyl; lower alkyl such as methyl, isopropyl or tert-butyl; lower alkylthio; xe2x80x94SCF3; xe2x80x94CH2OH; xe2x80x94COO-lower alkyl or xe2x80x94CONH2; and R39 is hydrogen, lower alkyl, aryl or aryl-lower alkyl.
In a further embodiment the invention relates to the compounds of the formula I wherein:
R1 and R2 independently are hydrogen or lower alkyl or together form a valence bond;
R3 and R4 independently are hydrogen or lower alkyl;
X is  greater than Cxe2x95x90O,  greater than Cxe2x95x90S,  greater than Cxe2x95x90NR5 or  greater than SO2;
n is 0, 1, 2 or 3;
m is 0 or 1;
R5 is hydrogen, lower alkyl, aryl-lower alkyl, or xe2x80x94OR6;
wherein R6 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
A is 
xe2x80x83wherein
R7 is hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR11, xe2x80x94NR11R12, lower alkyl, aryl, xe2x80x94SCF3, xe2x80x94SR11, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OSO2R11, xe2x80x94CONR11R12, xe2x80x94CH2OR11, xe2x80x94CH2NR11R12, xe2x80x94OCOR11, xe2x80x94CO2R13, xe2x80x94OSO2CF3;
R8 and R9 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR11, xe2x80x94NR11R12, lower alkyl, aryl, xe2x80x94SCF3, xe2x80x94SR11, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OSO2R11, xe2x80x94CONR11R12, xe2x80x94CH2OR11, xe2x80x94CH2NR11R12, xe2x80x94OCOR11, xe2x80x94CO2R13, xe2x80x94OSO2CF3, or R8 and R9 together form a bridge xe2x80x94OCH2Oxe2x80x94;
R11 and R12 independently are hydrogen, xe2x80x94COR13, xe2x80x94SO2R13, lower alkyl or aryl;
R13 is hydrogen, lower alkyl, aryl-lower alkyl or aryl;
R10 is hydrogen, lower alkyl, aryl-lower alkyl or aryl;
B is 
or a valence bond; preferably 
R14 and R15 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94O(CH2)lCF3, xe2x80x94NO2, xe2x80x94OR16, xe2x80x94NR16R17, lower alkyl, aryl, xe2x80x94SCF3, xe2x80x94SR16, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR16R17, xe2x80x94(CH2)lCONR16R17, xe2x80x94O(CH2)lCONR16R17, xe2x80x94(CH2)lCOR16, xe2x80x94O(CH2)lCOR16, xe2x80x94(CH2)lOR16, xe2x80x94O(CH2)lR16, xe2x80x94(CH2)lNR16R17, xe2x80x94O(CH2)lNR16R17, xe2x80x94OCOR16, xe2x80x94CO2R18, xe2x80x94O(CH2)lCN, xe2x80x94O(CH2)lCl, or R14 and R15 together form a bridge xe2x80x94Oxe2x80x94CH2xe2x80x94Oxe2x80x94;
R14 and R15 preferably independently representing hydrogen, halogen, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OR16, xe2x80x94NR16R17, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94OSO2CF3, xe2x80x94CONR16R17, xe2x80x94CH2OR16, xe2x80x94CH2NR16R17, xe2x80x94OCOR16 or xe2x80x94CO2R18; or together forming a bridge xe2x80x94OCH2Oxe2x80x94;
l is 1, 2, 3 or 4;
R16 and R17 independently are hydrogen, xe2x80x94COR18, xe2x80x94SO2R18, lower alkyl, aryl, or R16 and R17 together form a cyclic alkyl bridge containing from 2 to 7 carbon atoms;
R18 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
W is xe2x80x94Nxe2x95x90 or xe2x80x94CR19xe2x95x90;
Y is xe2x80x94Nxe2x95x90 or xe2x80x94CR20xe2x95x90;
Z is xe2x80x94Nxe2x95x90 or xe2x80x94CR21xe2x95x90;
V is xe2x80x94Nxe2x95x90 or xe2x80x94CR22xe2x95x90;
Q is xe2x80x94NR23xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
wherein:
R19, R20, R21 and R22 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR24, xe2x80x94NR24R25, lower alkyl, aryl, aryl-lower alkyl, SCF3, xe2x80x94SR24, xe2x80x94CHF2, xe2x80x94OCHF2, OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR24R25, xe2x80x94CH2CONR24R25, xe2x80x94OCH2CONR24R25, xe2x80x94CH2OR24, xe2x80x94CH2NR24R25, xe2x80x94OCOR24 or xe2x80x94CO2R24, or R19 and R20, R20 and R21 or R21 and R22 together form a bridge xe2x80x94OCH2Oxe2x80x94;
R24 and R25 independently are hydrogen, xe2x80x94COR26, xe2x80x94SO2R26, lower alkyl, aryl or aryl-lower alkyl;
R26 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
R23 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
K is 
xe2x80x83wherein:
R3a, R3b, R4b and R4b independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR24a, xe2x80x94NR24aR25a, lower alkyl, aryl, aryl-lower alkyl, SCF3, xe2x80x94SR24a, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR24aR25a, xe2x80x94CH2CONR24aR25a, xe2x80x94OCH2CONR24aR25a, xe2x80x94CH2OR24a, xe2x80x94CH2NR24aR25a, xe2x80x94OCOR24a or xe2x80x94CO2R24a;
wherein R24a and R25a independently are hydrogen, xe2x80x94COR26a, xe2x80x94SO2R26a, lower alkyl, aryl or aryl-lower alkyl;
R26a is hydrogen, lower alkyl, aryl or aryl-lower alkyl; or
R3a and R3b, R4a and R4b or R3a and R4b together form a bridge xe2x80x94(CH2)1xe2x80x94, wherein
i is 1, 2, 3 or 4;
a, b, c and d independently are 0, 1, 2, 3 or 4;
e, f, p and q independently are 0 or 1;
L and M independently are
xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94CHxe2x95x90CHxe2x80x94, xe2x80x94Cxe2x89xa1Cxe2x80x94, xe2x80x94NR5axe2x80x94, xe2x80x94COOxe2x80x94, xe2x80x94CONR5axe2x80x94, xe2x80x94NR5aCOxe2x80x94, xe2x80x94SOxe2x80x94, xe2x80x94SO2xe2x80x94, xe2x80x94OSO2xe2x80x94, xe2x80x94SO2xe2x80x94NR5axe2x80x94, xe2x80x94NR5aSO2xe2x80x94, xe2x80x94NR5aCONR5bxe2x80x94, xe2x80x94NR5aCSNR5bxe2x80x94, xe2x80x94OCONR5b or xe2x80x94NR5aC(O)Oxe2x80x94
wherein R5a and R5b independently are hydrogen, lower alkyl, xe2x80x94(CH2)kxe2x80x94OH, xe2x80x94(CH2)kxe2x80x94NR6aR6b, aryl or aryl-lower alkyl;
wherein k is 2, 3 or 4;
R6a and R6b independently are hydrogen, lower alkyl or aryl-lower alkyl;
K preferably representing 
D is hydrogen or 
preferably hydrogen, 
xe2x80x83wherein:
r and s independently are 1 or 2;
E, F and G independently are xe2x80x94CHR38xe2x80x94,  greater than Cxe2x95x90O,  greater than NR39, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
Yxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR32xe2x95x90;
Zxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR33xe2x95x90;
Vxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR34xe2x95x90;
Wxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR35xe2x95x90;
Qxe2x80x2 is xe2x80x94NR36xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
wherein
R27, R28, R32, R33, R34 and R35 are independently hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94O(CH2)yCF3, xe2x80x94NO2, xe2x80x94OR29, xe2x80x94NR29R30, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR29, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2R29, xe2x80x94OSO2CF3, xe2x80x94CONR29R30, xe2x80x94(CH2)yCONR29R30, xe2x80x94O(CH2)yCONR29R30, xe2x80x94(CH2)yOR29, xe2x80x94(CH2)yNR29R3, xe2x80x94OCOR29, xe2x80x94CO2R29; or R27 and R28, R32 and R33, R33 and R34 or R34 and R35 together form a bridge xe2x80x94OCH2Oxe2x80x94;
R27 and R28 preferably independently representing hydrogen, halogen, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94OR29, lower alkyl, aryl or aryl-lower alkyl, or together forming a bridge xe2x80x94OCH2Oxe2x80x94;
y is 1, 2, 3 or 4;
R29 and R30 independently are hydrogen, xe2x80x94COR31, xe2x80x94SO2R31, lower alkyl, aryl or aryl-lower alkyl;
R31 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
R36 and R39 independently are hydrogen, lower alkyl, aryl or aryl-lower alkyl;
R38 is hydrogen, xe2x80x94OR40, xe2x80x94NR40R41, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR40, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94CONR40R41, xe2x80x94(CH2)xCONR40R41, xe2x80x94O(CH2)xCONR40R41, xe2x80x94(CH2)xOR40, xe2x80x94(CH2)xNR40R41, xe2x80x94OCOR40 or xe2x80x94CO2R40;
x is 1, 2, 3 or 4;
R40 and R41 independently are hydrogen, xe2x80x94COR42, xe2x80x94SO2R42, lower alkyl, aryl or aryl-lower alkyl; and
R42 is hydrogen, lower alkyl, aryl or aryl-lower alkyl.
In a further embodiment the invention relates to the compounds of the formula I wherein:
R1 and R2 independently are hydrogen or lower alkyl or together form a valence bond;
R3 and R4 independently are hydrogen or lower alkyl;
n is 0, 1, 2 or 3;
m is 0 or 1;
X is  greater than Cxe2x95x90O,  greater than Cxe2x95x90S,  greater than Cxe2x95x90NR5 or  greater than SO2;
wherein R5 is hydrogen, lower alkyl, aryl-lower alkyl or xe2x80x94OR6;
wherein R6 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
A is 
xe2x80x83wherein:
R7 is hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR11, xe2x80x94NR11R12, lower alkyl, aryl, xe2x80x94SCF3, xe2x80x94SR11, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OSO2R11, xe2x80x94CONR11R12, xe2x80x94CH2OR11, xe2x80x94CH2NR11R12, xe2x80x94OCOR11, xe2x80x94CO2R13 or xe2x80x94OSO2CF3;
R8 and R9 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR11, xe2x80x94NR11R12, lower alkyl, aryl, xe2x80x94SCF3, xe2x80x94SR11, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OSO2R11, xe2x80x94CONR11R12, xe2x80x94CH2OR11, xe2x80x94CH2NR11R12, xe2x80x94OCOR11, xe2x80x94CO2R13 or xe2x80x94OSO2CF3, or R8 and R9 together form a bridge xe2x80x94OCH2Oxe2x80x94 or xe2x80x94OCH2CH2Oxe2x80x94;
wherein R11 and R12 independently are hydrogen, xe2x80x94COR13, xe2x80x94SO2R13, lower alkyl or aryl;
wherein R13 is hydrogen, lower alkyl, aryl-lower alkyl or aryl; and
R10 is hydrogen, lower alkyl, aryl-lower alkyl or aryl;
B is 
or a valence bond; preferably 
xe2x80x83wherein:
R14 and R15 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94O(CH2)lCF3, xe2x80x94NO2, xe2x80x94OR16, xe2x80x94NR16R17, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR16, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR16R17, xe2x80x94(CH2)lCONR16R17, xe2x80x94O(CH2)lCONR16R17, xe2x80x94(CH2)lCOR16, xe2x80x94(CH2)lCOR16, xe2x80x94(CH2)lOR16, xe2x80x94O(CH2)lOR16, xe2x80x94(CH2)lNR16R17, xe2x80x94O(CH2)lNR16R17, xe2x80x94OCOR16, xe2x80x94CO2R18, xe2x80x94O(CH2)lCO2R18, xe2x80x94O(CH2)lCN, xe2x80x94O(CH2)lCl, or R14 and R15 together form a bridge xe2x80x94OCH2Oxe2x80x94;
R14 and R15 preferably independently representing hydrogen, halogen, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OR16, xe2x80x94NR16R17, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94OSO2CF3, xe2x80x94CONR16R17, xe2x80x94CH2OR16, xe2x80x94CH2NR16R17, xe2x80x94OCOR16 or xe2x80x94CO2R18; or together forming a bridge xe2x80x94OCH2Oxe2x80x94;
wherein l is 1, 2, 3 or 4;
R16 and R17 independently are hydrogen, xe2x80x94COR18, xe2x80x94SO2R18, lower alkyl, aryl, or R16 and R17 together form a cyclic alkyl bridge containing from 2 to 7 carbon atoms;
wherein R18 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
W is xe2x80x94Nxe2x95x90 or xe2x80x94CR19xe2x95x90;
Y is xe2x80x94Nxe2x95x90 or xe2x80x94CR20xe2x95x90;
Z is xe2x80x94Nxe2x95x90 or xe2x80x94CR21xe2x95x90;
V is xe2x80x94Nxe2x95x90 or xe2x80x94CR22xe2x95x90; and
Q is xe2x80x94NR23xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
wherein:
R19, R20, R2 and R22 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR24, xe2x80x94NR24R25, lower alkyl, aryl, aryl-lower alkyl, SCF3, xe2x80x94SR24, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR24R25, xe2x80x94CH2CONR24R25, xe2x80x94OCH2CONR24R25, xe2x80x94CH2OR24, xe2x80x94CH2NR24R25, xe2x80x94OCOR24 or xe2x80x94CO2R24, or R19 and R20, R20 and R21 or R2 and R22 together form a bridge xe2x80x94OCH2Oxe2x80x94;
wherein R24 and R25 independently are hydrogen, xe2x80x94COR26, xe2x80x94SO2R26, lower alkyl, aryl or aryl-lower alkyl;
wherein R26 is hydrogen, lower alkyl, aryl or aryl-lower alkyl; and
R23 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
K is 
xe2x80x83wherein:
R3a, R3b, R4a and R4b independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCH2CF3, xe2x80x94NO2, xe2x80x94OR24a, xe2x80x94NR24aR25a, lower alkyl, aryl, aryl-lower alkyl, SCF3, SR24a, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2CF3, xe2x80x94CONR24aR25a, xe2x80x94CH2CONR24aR25a, xe2x80x94OCH2CONR24aR25a, xe2x80x94CH2OR24a, xe2x80x94CH2NR24aR25a, xe2x80x94OCOR24a or xe2x80x94CO2R24a;
wherein R24a and R25a independently are hydrogen, xe2x80x94COR26a, xe2x80x94SO2R26a, lower alkyl, aryl or aryl-lower alkyl;
wherein R26a is hydrogen, lower alkyl, aryl or aryl-lower alkyl; or
R3a and R3b, R4a and R4b or R3a and R4b together form a bridge xe2x80x94(CH2)ixe2x80x94;
wherein i is 1, 2, 3 or 4;
a, b, c and d independently are 0, 1, 2, 3 or 4;
e, f and p independently are 0 or 1;
q is 0,1 or 2; and
L and M independently are
xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94CHxe2x95x90CHxe2x80x94, xe2x80x94Cxe2x89xa1Cxe2x80x94, xe2x80x94NR5axe2x80x94, xe2x80x94COxe2x80x94, xe2x80x94OCOxe2x80x94, xe2x80x94COOxe2x80x94, xe2x80x94CONR5axe2x80x94, NR5aCOxe2x80x94, xe2x80x94SOxe2x80x94, xe2x80x94SO2xe2x80x94, xe2x80x94OSO2xe2x80x94, xe2x80x94SO2xe2x80x94NR5axe2x80x94, xe2x80x94NR5aSO2xe2x80x94, xe2x80x94NR5aCONR5bxe2x80x94, xe2x80x94NR5aCSNR5bxe2x80x94, xe2x80x94OCONR5bxe2x80x94 or xe2x80x94NR5aC(O)Oxe2x80x94;
wherein R5a and R5b independently are hydrogen, lower alkyl, xe2x80x94(CH2)kxe2x80x94OH, xe2x80x94(CH2)kxe2x80x94NR6aR6bxe2x80x94, aryl or aryl-lower alkyl;
wherein k is 2, 3 or 4; and
R6a and R6b independently are hydrogen, lower alkyl or aryl-lower alkyl;
K preferably representing 
D is hydrogen, 
preferably hydrogen 
xe2x80x83wherein:
r and s independently are 0, 1 or 2;
E, F and G independently are xe2x80x94CHR38xe2x80x94,  greater than Cxe2x95x90O,  greater than NR39, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
Fxe2x80x2 is  greater than CR38xe2x80x94 or  greater than Nxe2x80x94;
Yxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR32xe2x95x90;
Zxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR33xe2x95x90;
Vxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR34xe2x95x90;
Wxe2x80x2 is xe2x80x94Nxe2x95x90 or xe2x80x94CR35xe2x95x90; and
Qxe2x80x2 is xe2x80x94NR36xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94;
wherein:
R27, R28, R32, R33, R34 and R35 are independently hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94O(CH2)yCF3, xe2x80x94NO2, xe2x80x94OR29, xe2x80x94NR29R39, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR29, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94OSO2R29, xe2x80x94OSO2CF3, xe2x80x94CONR29R30, xe2x80x94(CH2)yCONR29R30, xe2x80x94O(CH2)yCONR29R30, xe2x80x94(CH2)yOR29, xe2x80x94(CH2)yNR29R30, xe2x80x94OCOR29 or xe2x80x94CO2R29; or
R27 and R28, R32 and R33, R33 and R34 or R34 and R35 together form a bridge xe2x80x94OCH2Oxe2x80x94;
R27 and R28 preferably independently representing hydrogen; halogen such as xe2x80x94Cl or xe2x80x94F; xe2x80x94CF3; xe2x80x94OCF3; xe2x80x94OCHF2; xe2x80x94OCH2CF3; xe2x80x94OR29 wherein R29 is hydrogen or lower alkyl; lower alkyl such as methyl, isopropyl or tert-butyl; lower alkylthio; xe2x80x94SCF3; xe2x80x94CH2OH; xe2x80x94COO-lower alkyl; aryl or xe2x80x94CONH2; or together forming a bridge xe2x80x94OCH2Oxe2x80x94;
wherein y is 1, 2, 3 or 4; and
R29 and R30 independently are hydrogen, xe2x80x94COR31, xe2x80x94SO2R31, lower alkyl, aryl or aryl-lower alkyl;
wherein R31 is hydrogen, lower alkyl, aryl or aryl-lower alkyl;
R36 and R39 independently are hydrogen, lower alkyl, aryl or aryl-lower alkyl; and
R38 is hydrogen, xe2x80x94OR40, xe2x80x94NR40R41, lower alkyl, aryl, aryl-lower alkyl, xe2x80x94SCF3, xe2x80x94SR40, xe2x80x94CHF2, xe2x80x94OCHF2, xe2x80x94OCF2CHF2, xe2x80x94CONR40R41, xe2x80x94(CH2)xCONR40R41, xe2x80x94O(CH2)xCONR40R41, xe2x80x94(CH2)xOR40, xe2x80x94(CH2))xNR40R41, xe2x80x94OCOR40 or xe2x80x94CO2R40;
wherein x is 1, 2, 3 or 4;
R40 and R41 independently are hydrogen, xe2x80x94COR42, xe2x80x94SO2R42, lower alkyl, aryl or aryl-tower alkyl; and
wherein R42 is hydrogen, lower alkyl, aryl or aryl-lower alkyl.
Examples of specific compounds represented by the above general formula V are the following: 
The most preferred specific compounds represented by the above general formula III are the following: 
The most preferred specific compounds represented by the above general formula IV are the following: 
Preferred specific compounds represented by the formulae VI and VII are the following: 
The most preferred specific compounds of formula I wherein A is a heterocyclic and/or bicyclic moiety are the following: 
Especially preferred according to the present invention are the following compounds which show a particularly high affinity to the human glucagon receptor: 
The compounds of the present invention may have one or more asymmetric centres and it is intended that any optical isomers, as separated, pure or partially purified optical isomers or racemic mixtures thereof are included in the scope of the invention.
Furthermore, one or more carbon-carbon or carbon-nitrogen double bonds may be present in the compounds which brings about geometric isomers. It is intended that any geometric isomers, as separated, pure or partially purified geometric isomers or mixtures thereof are included in the scope of the invention.
Furthermore, the compounds of the present invention may exist in different tautomeric forms, eg the following tautomeric forms: 
It is intended that any tautomeric forms which the compounds are able to form are included in the scope of the present invention.
Owing to their efficacy in antagonizing the glucagon receptor the present compounds may be suitable for the treatment and/or prevention of any glucagon-mediated conditions and diseases.
Accordingly, the present compounds may be applicable for the treatment of hyperglycemia associated with diabetes of any cause or associated with other diseases and conditions, eg impaired glucose tolerance, insulin resistance syndromes, syndrome X, type I diabetes, type II diabetes, hyperlipidemia, dyslipidemia, hypertriglyceridemia, glucagonomas, acute pancreatitis, cardiovascular diseases, cardiac hypertrophy, gastrointestinal disorders, diabetes as a consequence of obesity etc. Furthermore, they may be applicable as diagnostic agents for identifying patients having a defect in the glucagon receptor, as a therapy to increase gastric acid secretions, to reverse intestinal hypomobility due to glucagon administration, to reverse catabolism and nitrogen loss in states of negative nitrogen balance and protein wasting including all causes of type I and type II diabetes, fasting, AIDS, cancer, anorexia, aging and other conditions, for the treatment of any of the above conditions or diseases post-operative or during surgery and for decreasing saitety and increasing energy intake. Thus, in a further aspect the present invention relates to a pharmaceutical composition comprising, as an active ingredient, at least one compound according to the present invention together with one or more pharmaceutically acceptable carriers or excipients.
The present invention furthermore relates to methods of treating type I or type II diabetes or hyperglycemia which methods comprise administering to a subject in need thereof an effective amount of a compound according to the invention.
Moreover, the present invention relates to a method of lowering blood glucose in a mammal, comprising administering to said mammal an effective amount of a compound according to the invention.
The present invention is also concerned with the use of a compound according to the invention for the manufacture of a medicament for treating type I or type II diabetes or hyperglycemia, or for lowering blood glucose in a mammal.
The compounds according to the invention, which may also be referred to as an active ingredient, may be administered for therapy by any suitable route including oral, rectal, nasal, pulmonal, topical (including buccal and sublingual), transdermal, vaginal and parenteral (including subcutaneous, intramuscular, intravenous and intradermal), the oral route being preferred. It will be appreciated that the preferred route will vary with the condition and age of the recipient, the nature of the condition to be treated, and the chosen active ingredient.
The compounds of the invention are effective over a wide dosage range. A typical dosage is in the range of from 0.05 to about 1000 mg, preferably of from about 0.1 to about 500 mg, such as of from about 0.5 mg to about 250 mg for administration one or more times per day such as 1 to 3 times per day. It should be understood that the exact dosage will depend upon the frequency and mode of administration, the sex, age, weight and general condition of the subject treated, the nature and severity of the condition treated and any concomitant diseases to be treated as well as other factors evident to those skilled in the art.
The formulations may conveniently be presented in unit dosage form by methods known to those skilled in the art.
For parenteral routes, such as intravenous, intrathecal, intramuscular and similar administration, typically doses are on the order of about xc2xd the dose employed for oral administration.
The compounds of this invention are generally utilized as the free substance or as a pharmaceutically acceptable salt thereof. One example is an acid addition salt of a compound having the utility of a free base. When a compound of formula I contains a free base such salts are prepared in a conventional manner by treating a solution or suspension of a free base of formula I with a chemical equivalent of a pharmaceutically acceptable acid, for example, inorganic and organic acids, for example: maleic, fumaric, benzoic, ascorbic, pamoic, succinic, bismethylene salicylic, methanesulfonic, ethanedisulfonic, acetic, oxalic, propionic, tartaric, salicylic, citric, pyruvic, gluconic, lactic, malic, mandelic, cinnamic, citraconic, aspartic, stearic, palmitic, EDTA, glycolic, p-aminobenzoic, glutamic, benzenesulfonic, p-toluensulfonic, hydrochloric, hydrobromic, sulfuric, phosphoric or nitric acids. Physiologically acceptable salts of a compound with a hydroxy group include the anion of said compound in combination with a suitable cation such as sodium or ammonium ion.
The compounds of the invention may be administered alone or in combination with pharmaceutically acceptable carriers, in either single or multiple doses.
For parenteral administration, solutions of the novel compounds of formula I in sterile aqueous solution, aqueous propylene glycol or sesame or peanut oil may be employed. Such aqueous solutions should be suitable buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. The aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. The sterile aqueous media employed are all readily available by standard techniques known to those skilled in the art. Suitable pharmaceutical carriers include inert solid diluents or fillers, sterile aqueous solution and various organic solvents. Examples of solid carriers are lactose, terra alba, sucrose, cyclodextrin, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid or lower alkyl ethers of cellulose. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids, fatty acid amines, polyoxyethylene or water. Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax. The pharmaceutical compositions formed by combining the novel compounds of formula I and the pharmaceutically acceptable carriers are then readily administered in a variety of dosage forms suitable for the disclosed routes of administration. The formulations may conveniently be presented in unit dosage form by methods known in the art of pharmacy.
Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules or tablets, each containing a predetermined amount of the active ingredient, and which may include a suitable excipient. These formulations may be in the form of powder or granules, as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion.
If a solid carrier is used for oral administration, the preparation may be tabletted, placed in a hard gelatin capsule in powder or pellet form or it can be in the form of a troche or lozenge. The amount of solid carrier will vary widely but will usually be from about 25 mg to about 1 g.
If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.
A typical tablet which may be prepared by conventional tabletting techniques may contain:
For nasal administration, the preparation may contain a compound of formula I dissolved or suspended in a liquid carrier, in particular an aqueous carrier, for aerosol application. The carrier may contain additives such as solubilizing agents, e.g. propylene glycol, surfactants, absorption enhancers such as lecithin (phosphatidylcholine) or cyclodextrin, or preservatives such as parabenes.
Optionally, the pharmaceutical composition of the invention may comprise a compound of formula I combined with one or more other pharmacologically active compounds, e.g. an antidiabetic or other pharmacologically active material, including compounds for the treatment and/or prophylaxis of insulin resistance and diseases wherein insulin resistance is the patophysiological mechanism. Suitable antidiabetics comprise insulin, GLP-1 derivatives such as those disclosed in WO 98/08871 (Novo Nordisk A/S) which is incorporated herein by reference as well as orally active hypoglycaemic agents such as sulphonylureas, e.g. glibenclamide and glipizide; biguanides, e.g. metformin; benzoic acid derivatives, e.g. repaglinide; and thiazolidinediones, e.g. troglitazone and ciglitazone, as well as PPAR and RXR agonists.
In the following section binding assays as well as functional assays useful for evaluating the efficacy of the compounds of the invention are described.
Binding of compounds to the glucagon receptor was determined in a competition binding assay using the cloned human glucagon receptor.
In the screening setup, antagonism was determined as the ability of the compounds to inhibit the amount of cAMP formed in the presence of 5 nM glucagon.
For full characterization, antagonism was determined in a functional assay, measured as the ability of the compounds to right-shift the glucagon dose-response curve. Using at least 3 different antagonist concentrations, the Ki was calculated from a Schild plot. Receptor binding was assayed using cloned human receptor (Lok et al, Gene 140, 203-209 (1994)). The receptor inserted in the pLJ6xe2x80x2 expression vector using EcoRI/SSt1 restriction sites (Lok et al) was expressed in a baby hamster kidney cell line (A3 BHK 570-25). Clones were selected in the presence of 0.5 mg/ml G-418 and were shown to be stable for more than 40 passages. The Kd was shown to be 0.1 nM.
Plasma membranes were prepared by growing cells to confluence, detaching them from the surface and resuspending the cells in cold buffer (10 mM tris/HCl), pH 7.4 containing 30 mM NaCl, 1 mM dithiothreitol, 5 mg/l leupeptin (Sigma), 5 mg/l pepstatin (Sigma), 100 mg/l bacitracin (Sigma) and 15 mg/l recombinant aprotinin (Novo Nordisk)), homogenization by two 10-s bursts using a Polytron PT 10-35 homogenizer (Kinematica), and centrifugation upon a layer of 41 w/v % sucrose at 95.000*g for 75 min. The white band located between the two layers was diluted in buffer and centrifuged at 40.000*g for 45 min. The precipitate containing the plasma membranes was suspended in buffer and stored at xe2x88x9280xc2x0 C. until required.
Glucagon was iodinated according to the chloramine T method (Hunter and Greenwood, Nature 194, 495 (1962)) and purified using anion exchange chromatography (Jxc3x8rgensen et al, Hormone and Metab. Res. 4, 223-224 (1972). The specific activity was 460 xcexcCi/xcexcg on day of iodination. Tracer was stored at xe2x88x9218xc2x0 C. in aliquots and were used immediately after thawing.
Binding assays were carried out in triplicate in filter microtiter plates (MADV N65, Millipore). The buffer used in this assay was 25 mM HEPES pH 7.4 containing 0.1% human serum albumin (Sigma, grade V). Glucagon was dissolved in 0.05 M HCl, added equal amounts(w/w) of HSA and freeze-dried. On the day of use, it was dissolved in water and diluted in buffer to the desired concentrations.
175 xcexcl of sample (glucagon or test compounds) was added to each well. Tracer (50.000 cpm) was diluted in buffer and 15 xcexcl was added to each well. 0.5 xcexcg freshly thawed plasma membrane protein diluted in buffer was then added in 15 xcexcl to each well. Plates were incubated at 25xc2x0 C. for 2 hours. Non specific binding was determined with 10xe2x88x926 M glucagon. Bound and unbound tracer were then separated by vacuum filtration (Millipore vacuum manifold). The plates were washed once with 150 xcexcl buffer/well. The plates were air dried for a couple of hours, whereafter filters were separated from the plates using a Millipore Puncher. The filters were counted in a xcex3 counter.
The functional assay was carried out in 96 well microtiter plates (tissue culture plates, Nunc). The resulting buffer concentrations in the assay were 50 mM tris/HCl, 1 mM EGTA, 1.5 mM MgSO4, 1.7 mM ATP, 20 xcexcM GTP, 2 mM IBMX, 0.02% tween-20 and 0.1% HSA. pH was 7.4 Glucagon and proposed antagonist were added in 35 xcexcl diluted in 50 mM tris/HCl, 1 mM EGTA, 1.85 mM MgSO4, 0.0222% tween-20 and 0.111% HSA, pH 7.4. 20 xcexcl of 50 mM tris/HCl, 1 mM EGTA, 1.5 mM MgSO4, 11.8 mM ATP, 0.14 mM GTP, 14 mM iso-buthyl-methylxanthine (IBMX) and 0.1% HSA, pH 7.4 was added. GTP was dissolved immediately before the assay.
50 xcexcl containing 5 xcexcg plasma membrane protein was added in a tris/HCl, EGTA, MgSO4, HSA buffer (the actual concentrations were dependent upon the concentration of protein in the stored plasma membranes).
The total assay volume was 140 xcexcl. The assay was incubated for 2 hours at 37xc2x0 C. with continuous shaking. Reaction was terminated by addition of 25 xcexcl 0.5 N HCl. cAMP was measured by the use of a scintillation proximity kit (Amersham).
Receptor binding was assayed using the cloned human receptor (Lok et al, Gene 140, 203-209 (1994)). The receptor inserted in the pLJ6xe2x80x2 expression vector using EcoRI/SSt1 restriction sites (Lok et al) was expressed in a baby hamster kidney cell line (A3 BHK 570-25). Clones were selected in the presence of 0.5 mg/ml G-418 and were shown to be stable for more than 40 passages. The Kd was shown to be 0.1 nM.
Plasma membranes were prepared by growing cells to confluence, detaching them from the surface and resuspending the cells in cold buffer (10 mM tris/HCl), pH 7.4 containing 30 mM NaCl, 1 mM dithiothreitol, 5 mg/l leupeptin Sigma), 5 mg/l pepstatin (Sigma), 100 mg/l bacitracin (Sigma) and 15 mg/l recombinant aprotinin (Novo Nordisk)), homogenization by two 10-s bursts using a Polytron PT 10-35 homogenizer (Kinematica), and centrifugation. The homogenate was resuspended and centrifuged again. The final precipitate containing the plasma membranes was suspended in buffer and stored at xe2x88x9280xc2x0 C. until required.
Binding assays were carried out in duplicate in polypropylene tubes or microtiter plates. The buffer used in this assay was 25 mM HEPES pH 7.4 containing 0.1% bovine serum albumin (Sigma, fraction V). Sample (glucagon (Bachem CA) or test compounds) was added to each tube or well. Tracer (xe2x88x9225000 cpm) was diluted in buffer and was added to each tube or well. 0.5 xcexcg freshly thawed plasma membrane protein diluted in buffer was then added in aliquots to each tube or well. Tubes or plates were incubated at 37xc2x0 C. for 1 hour. Non specific binding was determined with 10xe2x88x927 M glucagon. Bound and unbound tracer were then separated by vacuum filtration (Brandel). The tubes or wells were washed twice with buffer. The filters or plates were counted in a gamma counter.
The functional assay determined the ability of the compounds to antagonize glucagonstimulated formation of cAMP in a whole-cell assay. The assay was carried out in borosilicate 5 glass 12xc3x9775 tubes. The buffer concentrations in the assay were 10 mM HEPES, 1 mM EGTA, 1.4 mM MgCl2, 0.1 mM IBMX, 30 mM NaCl, 4.7 mM KCl, 2.5 mM NaH2PO4, 3mM glucose and 0.2% BSA. The pH was 7.4. Loose whole cells (0.5 ml, 106/ml) were pretreated with various concentrations of compounds for 10 min at 37xc2x0 C., then challenged with glucagon for 20 min. Some aliquots (500 xcexcL) of cells were treated with test compounds (55 xcexcL) alone to test for agonist activity. The reactions were terminated by centrifugation, followed by cell lysis with the addition of 500 xcexcl 0.1% HCl. Cellular debris was pelleted and the supernatant containing cAMP evaporated to dryness. cAMP was measured by the use of an RIA kit (NEN, NEK-033). Some assays were carried out utilizing the adenylate cyclase FlashPlate system from NEN.
The following synthesis protocols refer to intermediate compounds and final products identified in the specification and in the synthetic schemes. The preparation of the compounds of the present invention is described in detail using the following examples, but the chemical reactions described are disclosed in terms of their general applicability to the preparation of the glucagon antagonists of the invention. Occasionally, the reaction may not be applicable as described to each compound included within the disclosed scope of the invention. The compounds for which this occurs will be readily recognized by those skilled in the art. In all such cases, either the reactions can be successfully performed by conventional modifications known to those skilled in the art, that is, by appropriate protection of interfering groups, by changing to other conventional reagents, or by routine modification of reaction conditions. Alternatively, other reactions disclosed herein or otherwise conventional will be applicable to the preparation of the corresponding compounds of the invention. In all preparative methods, all starting materials are known or readily preparable from known starting materials. All temperatures are set forth in degrees Celsius and unless otherwise indicated, all parts and percentages are by weight when referring to yields and all parts are by volume when referring to solvents and eluents.
The compounds of general formula I may be prepared according to one embodiment of the invention, the alkylidene hydrazides of general formula II, as indicated in Scheme I, that is, by converting an ester of a carboxylic acid, for example, an aromatic acid to a hydrazide derivative and further reacting that product compound with a substituted aldehyde or ketone to yield a substituted alkylidene hydrazide. 
wherein A, B, K, D, m, n and R4 are as defined for formula I and R3 is lower alkyl.
The reaction is known (Org. Syn., Coll. Vol. II, A. H. Blatt, ed., John Wiley and Sons, New York, 1943, p. 85; Org. Syn., Coll. Vol. IV, N. Rabjohn, ed., John Wiley and Sons, New York, 1963, p. 819) and is generally performed by stirring the corresponding ester (either methyl, ethyl or other lower alkyl ester) with 2-10 molar excess of hydrazine in the presence of a solvent such as ethyl alcohol, methyl alcohol, isopropyl or tert-butyl alcohol or tetrahydrofuran, dioxane, DMSO, ethylene glycol, ethylene glycol dimethyl ester, benzene, toluene or a mixture of the above solvents or, in the absence of a solvent where excess of hydrazine acts as a solvent. The reactions are performed between 0xc2x0 C. to 130xc2x0 C., preferably between 20xc2x0 C. to 10xc2x0 C., most preferably at or about the reflux temperature of the solvent. The reactions are preferably conducted under an inert atmosphere such as N2 or Ar. When the reaction is complete as judged by disappearance of the starting ester by TLC or HPLC, the solvent may be removed by concentration at atmospheric or reduced pressure.
The product can be further purified by either recrystallization from a solvent such as ethyl alcohol, methyl alcohol, isopropyl alcohol, toluene, xylene, hexane, tetrahydrofuran, diethyl ether, dibutyl ether, water or a mixture of two or more of the above. Alternatively, the product can be purified by column chromatography using dichloromethane/methanol or chloroform/methanol or isopropyl alcohol as eluent. The corresponding fractions are concentrated either at atmospheric pressure or in vacuo to provide the pure aroyl hydrazide.
The methyl or ethyl ester of the corresponding aromatic acid, such as for example a substituted benzoic acid ester, is dissolved in ethanol and hydrazine (5 eq) is added. The reaction is refluxed overnight under nitrogen. Upon cooling the substituted hydrazide derivative usually precipitates. After filtration the product is usually recrystallized from hot methanol, ethanol or isopropyl alcohol. In cases where the hydrazide does not precipitate, the reaction is concentrated under vacuo and chromatographed over silica gel using dichloromethane/methanol as the eluent. Specific examples illustrating the preparation of aromatic hydrazides are provided below.
To a sample of ethyl 5-hydroxyindole-2-carboxylate (5 g, 24 mmol), dissolved in ethanol (250 mL) was added hydrazine (4 mL, 121 mmol). The reaction was refluxed overnight under nitrogen. Upon cooling the reaction vessel, the desired product crystallized. The white solid was isolated by filtration. Recrystallization from hot ethanol gave the 5-hydroxyindole-3-carboxylic acid hydrazide in 85% yield. 
1H NMR (DMSO-d6): xcex4 4.38 (s, 2H); 6.62 (dd, 1H); 6.76 (dd, 2H); 7.13 (d, 1H); 8.70 (s, 1H); 9.57 (s, 1H); 11.21 (s, 1H); MS (FAB): m/z 192 (M+H)+.
To a sample of methyl 3-chloro-4-hydroxybenzoate (2 g) dissolved in ethanol (50 mL) was added hydrazine (1.8 mL). The reaction was refluxed overnight under nitrogen. Upon cooling the reaction vessel, the desired product crystallized out of solution. The white solid was isolated by filtration. Recrystallization from hot ethanol gave the 3-chloro-4-hydroxybenzoic acid hydrazide in 60% yield. 
1H NMR (DMSO-d6): xcex4 4.49 (broad s, 2H), 7.05 (dd, 1H), 7.71 (dd, 1H), 7.89 (d, 1H), 9.669 (s, 1H), 10.72 (broad s, 1H).
By use of the above methodology, other hydrazides useful as intermediates in preparing the compounds of the invention are prepared, for example: 
3-Bromo-4-hydroxybenzoic Acid Hydrazide
1H NMR (DMSO-d6): xcex4 9.95 (s, 1H), 9.65 (d, 1H), 9.61 (broad s, 1H), 6.95 (d, 1H), 4.40 (broad s, 2H); MS m/z 233.1. 
3-Nitro-4-hydroxybenzoic Acid Hydrazide
1H NMR (DMSO-d6): xcex4 9.28 (broad s, 1H), 8.28 (s, 1H), 7.52 (d, 1H), 6.41 (d, 1H). MS m/z 198. 
3-Fluoro-4-hydroxybenzoic Acid Hydrazide
1H NMR (DMSO-d6): xcex4 9.45 (broad s, 1H), 7.5 (d, 1H), 7.43 (d, 1H), 6.85 (t, 1H), 5.55 (broad s, 3H).

Step A:
4-amino-2-chlorobenzoic acid (10 g, 58 mmol) was dissolved in H2SO4 (12 N, 120 mL) with heating. After cooling the solution in an ice-bath aqueous NaNO2 (2.5 M, 25 mL) was added dropwise such that the internal temperature remained at 5xc2x0 C. Urea was added to the mixture for after stirring for 15 minutes to destroy excess NaNO2 (monitored by starch iodine test). CuSO4 (100-200 mg) was added and the mixture was heated to 90xc2x0 C. until evolution of gas stopped. After cooling, the mixture was extracted with ethyl ether (3xc3x97). The combined organic fractions were extracted with 3N NaOH (3xc3x97). The combined aqueous layer was acidified with conc. HCl and the product was extracted with ethyl ether (3xc3x97). The organic fractions were washed with water, brine, and dried over MgSO4. The crude product was introduced into a silica gel column and eluted with ethyl acetate/hexane (1/1) to afford 2-chloro-4-hydroxybenzoic acid.
1H NMR (DMSO-D6): xcex4 6.97 (dd, 1H), 7.05 (d, 1H), 7.95 (d, 1H), 10.90 (brd s, 1H).
Step B:
To a solution 2-chloro-4-hydroxybenzoic acid in anhydrous methanol was added thionyl chloride (1.5 eq). After stirring the solution at room temperature for 16 hours, the solvent was evaporated. The residue was taken up in ethyl acetate and washed with saturated aqueous sodium bicarbonate, water, brine, and dried over MgSO4 and concentrated in vacuo to give methyl 2-chloro-4-hydroxybenzoate.
Step C:
To a solution of methyl 2-chloro-4-hydroxybenzoate (13.6 g, 73.1 mmol) in acetic acid (300 mL) was added N-chlorosuccinimide (9.8 g, 73.7 mmol). The solution was refluxed for 24 h and the solvent was evaporated under vacuo. The residue was taken up in chloroform, washed with water, brine, dried over magnesium sulfate, filtered and concentrated. Methyl 2,3-dichloro-4-hydroxybenzoate precipitated out of ethyl acetate. Chromatography of the residue using ethyl acetate/hexane (1/9 to 3/7) afforded methyl 2,5-dichloro-4-hydroxybenzoate (1.4 g, 60%) as well as an additional batch of methyl 2,3-dichloro-4-hydroxybenzoate isomer (total of 8.4 g, 10%).
Methyl 2,3-dichloro-4-hydroxybenzoate:
1H NMR (DMSO-D6) xcex4 3.81 (s, 3H), 7.02 (d, 1H), 7.70 (d 1H), 11.52 (s, 1H); MS (APCI): 221, 223.
Methyl 2,5-dichloro-4-hydroxybenzoate:
1H NMR (CDCl3): xcex4 3.90 (s, 3H), 6.00 (s, 1H), 7.14 (s, 1H), 7.27 (s, 1H), 7.96 (s, 1H); MS (APCI): 221.9.
Step D:
The title compound was prepared according to the general procedure for the synthesis of precursor hydrazides Axe2x80x94(Cxe2x95x90O)xe2x80x94NHNH2.
1H NMR (DMSO-D6): xcex4 6.82 (dd, 1H), 6.90 (d, 1H), 7.79 (d, 1H, 10.68 (brd s, 1H).
The 2,3-dichloro-4-hydroxybenzoic Acid Hydrazide was prepared from the methyl 2,3-dichloro-4-hydroxybenzoate above according to the general procedure for the synthesis of precursor hydrazides Axe2x80x94(Cxe2x95x90O)xe2x80x94NHNH2 with the exception that pentanol was the solvent of choice. The product was purified via silica gel column chromatography using CH2Cl2/MeOH (95/5 to 80/20), yield=50%.
2,5-dichloro-4-hydroxybenzoic acid hydrazide was prepared in a similar way starting from 2,5-dichloro-4-hydroxybenzoate.
2,3-Dichloro-4-hydroxybenzoic Acid Hydrazide:
1H NMR (DMSO-D6) xcex4 4.41 (brd s, 2H), 6.99 (1, 1H), 7.37 (s, 1H), 9.46 (s, 1H), 11.04 (s, 1H).
2,5-Dichloro-4-hydroxybenzoic Acid Hydrazide:
1H NMR (DMSO-D6) xcex4 4.48 (brd s, 3H), 6.92 (d, 2H), 7.18 (d, 2H), 9.45 (brd s, 1H).

Step A:
A mixture of 2,3-difluoro-4-cyanophenol (1 g, 6.45 mmol) in water (8 mL), H2SO4 (8 mL), and acetic acid (8 mL) was refluxed for 48 hours. The solvents were removed by rotary evaporation to give a slurry which was poured onto ice. The product precipitated out of solution and filtered. The solid was washed with water and dried to give 2,3-difluoro-4-hydroxybenzoic acid (800 mg, 71%).
1H NMR (DMSO-D6): xcex4 6.87 (t, 1H), 7.60 (t, 1H), 11.28 (s, 1H), 12.53 (brd s, 1H).
Step B:
To the 2,3-difluoro-4-hydroxybenzoic acid (800 mg, 5.1 mmol) dissolved in anhydrous methanol (50 mL) was added thionyl chloride (0.55 mL, 7.3 mmol). After stirring the solution at room temperature for 16 hours, the solvent was evaporated. The residue was taken up in ethyl acetate and washed with saturated aqueous sodium bicarbonate, water, brine, and dried over MgSO4 to give methyl 2,3-difluoro-4-hydroxybenzoate (540 mg, 62%).
1H NMR (CDCl3): xcex4 3.92 (s, 3H), 6.34 (brd s, 1H), 6.82 (dt, 1H), 7.68 (dt, 1H).
Step C:
The 2,3-difluoro-4-hydroxybenzoic acid hydrazide was prepared from the methyl 2,3-difluoro-4-hydroxybenzoate above according to the general procedure for the synthesis of precursor hydrazides Axe2x80x94(Cxe2x95x90O)xe2x80x94NHNH2. The product was purified via silica gel column chromatography using CH2Cl2/MeOH (95/5 to 80/20) to afford the title compound.
1H NMR (DMSO-D6): xcex4 4.48 (s, 2H), 6.80 (m, 1H), 7.22 (m, 1H), 9.36 (s, 1H), 10.89 (s, 1H); MS (APCI): 189.

Step A:
Methyl-4-hydroxybenzoate (35.5 g, 0.233 mol) was dissolved in 200 mL of warm (65xc2x0 C.) acetic acid. A solution of iodine monochloride (37.8 g, 0.233 mol) in 50 mL of acetic acid was added slowly (40 minutes) to the methyl-4-hydroxybenzoate solution, while maintaining a temperature of 65xc2x0 C. and vigorous stirring. The product crystallizes from solution upon cooling to room temperature and standing overnight. The crystals were collected on a filter, washed with water, then dried under vacuum. Methyl-4-hydroxy-3-iodobenzoate was obtained as white crystals (28.6 g, 44%).
1H NMR (DMSO-D6): xcex4 3.79 (s, 3H), 6.95 (d, J=8.3, 1H), 7.81 (dd, J=8.3, 2.2, 1H), 8.22 (d, J=2.2, 1H); 13C NMR (DMSO-D6) xcex4 52.8, 85.2, 115.5, 123.0, 132.0, 141.0, 161.9, 165.6; MS (APCI, neg): 277.
Step B:
Methyl-4-hydroxy-3-iodobenzoate (2.00 g, 7.2 mmol) was dissolved into 5 mL of dry DMF. Copper(I) cyanide (0.72 g, 8.0 mmol) and a small crystal of sodium cyanide was added. The mixture was flushed with nitrogen, placed in an oil heating bath (100-110xc2x0 C.), and stirred overnight. TLC indicated nearly complete reaction. The mixture was cooled and the solids removed by filtration. The solids were extracted with DMF (3 mL). The filtrate and washings were taken up in 100 mL of ethyl acetate, then washed with 3 portions of saturated sodium chloride solution. The solids and aqueous washings were combined, and shaken with a mixture of 50 mL of ethyl acetate and a ferric chloride solution (4 g of hydrated ferric chloride in 7 mL of conc. hydrochloric acid). The ethyl acetate layers were combined, washed with brine containing sodium metabisulfite, dried over sodium sulfate, filtered, and the solvent removed in vacuo. The resulting solids were purified by flash chromatography on silica gel (20% ethyl acetate/hexane) to afford methyl-3-cyano-4-hydroxybenzoate, 0.93g (73%).
1H NMR (DMSO-D6): xcex4 3.79 (s, 3H), 7.07 (d, J=8.7, 1H), 8.02 (dd, J=8.7, 1.9, 1H), 8.10 (d, J=1.9, 1H).
Step C:
Methyl-3-cyano-4-hydroxybenzoate (2.71 g, 15.3 mmol) was dissolved in 50 mL of THF. The solution was chilled in an ice bath, and 2.0M potassium hydroxide (17 mL, 34 mmol) was added dropwise. The resulting mixture was stirred at room temperature overnight. TLC indicated complete reaction. The THF was removed by rotary evaporation. The aqueous residue was acidified with aqueous trifluoroacetic acid and purified by reverse-phase HPLC (C-18, 0.1% TFA in water and acetonitrile). 3-Cyano-4-hydroxybenzoic acid was obtained as a white powder (2.1 g, 84%) after lyophilization.
1H NMR (DMSO-D6): xcex4 7.09 (d, J=9.0, 1H), 8.00 (dd, J=9.0, 2.3, 1H), 8.07 (d, J=2.3, 1H) 12.50 (br s, 2H); MS (APCI, neg): 162. IR: 2252 cmxe2x88x921, CN.
Step D:
3-Cyano-4-hydroxybenzoic acid (1.88 g, 11.5 mmol) was dissolved in 20 mL of methylene chloride/DMF (1/1) and chilled in an ice-bath. Diisopropylethylamine (12 mL, 69 mmol), t-butyl carbazate (1.76 g, 13.3 mmol), and PyBroP (bromo-tris-pyrrolidino-phosphonium hexafluorophosphate, 6 g, 12.9 mmol) were added, and the mixture was stirred to form a clear solution. The solution stood in the refrigerator overnight. TLC indicated that the reaction was not complete, so additional diisopropylethylamine (22 mL, 127 mmol), t-butyl carbazate (0.85 g, 6.4 mmol) and PyBroP (3.0 g, 6.4 mmol) were added. After 8 more hours at 0xc2x0 C., the reaction was worked up as follows. The solution was reduced by rotary evaporation. The remaining DMF solution was diluted with 100 mL of ethyl acetate, and washed with several portions of 0.1 M HCl (until the wash remained acidic to litmus paper). The ethyl acetate layer was further washed with 3 portions of brine, dried over magnesium sulfate, filtered, and reduced to an oil in vacuo. The oil was purified by chromatography on silica gel (6:4 hexane:ethyl acetate) to afford tert-butyloxycarbonyl (3-cyano-4-hydroxy)benzoic acid hydrazide as a white solid (1.8 g, 56%).
1H NMR (DMSO-D6): xcex4 1.42 (s, 9H), 7.09 (d, J=8.7, 1H), 7.98 (m, 1H), 8.11 (br s, 1H), 8.92 (s, 1H), 10.15 (s, 1H), 11.73 (br s, 1H); MS (APCI, neg): 276; IR: 2232 cmxe2x88x921, CN.
Step E:
The Boc-hydrazide (1.8 g, 6.5 mmol) was suspended in 50 mL of chloroform and cooled in an ice-bath. Trifluoroacetic acid was added with stirring, and the resulting solution stood for 4 hours at 0xc2x0 C. TLC indicated complete reaction. Solvent and excess TFA were removed by rotary evaporation. The remaining oil was purified by reverse-phase liquid chromatography (Aquasil C-18 column, water/acetonitrile/0.1% TFA). The title compound was obtained as a white solid (0.24 g, 13%).
1H NMR (DMSO-D6): xcex4 7.16 (d, J=9.0, 1H), 8.00 (dd, J=1.5, 9.0, 1H), 8.14 (d, J=1.5, 1H), 10.47 (br s, 5H); MS (APCI, neg): 176.

Step A:
Silver nitrate (17 g, 0.1 mol) was dissolved in water (10 mL) and treated with 1 N NaOH (300 mL, 0.3 mol). The brown precipitate which was formed was stirred for 30 minutes and the supernatant was decanted. The brown silver oxide was washed with additional volumes of water (3xc3x97).
To the silver oxide above was added 1N NaOH (150 mL) and 4-hydroxynaphthaldehyde (1 g, 6 mmol)). The mixture was heated to 70xc2x0 C. for 10 minutes after which additional amounts of 4-hydroxynaphthaldehyde (5.5 g, 32 mmol) was added in portions. The mixture was kept at 80xc2x0 C. for 16 hours. TLC analysis indicated incomplete conversion. An additional portion of silver oxide was prepared as above and added to the reaction mixture. After heating the mixture for an additional 6 hours, the mixture was cooled and acidified with 1N HCl. The aqueous layer was extracted with ethyl acetate (3xc3x97) and upon concentration 4-hydroxynaphthoic acid precipitated (3.7 g, 60%) out of solution.
1H NMR (DMSO-D6): xcex4 6.69 (d, 1H), 7.28 (t, 1H), 7.39 (t, 1H), 7.93 (d, 1H), 8.03 (d, 1H), 8.82 (d, 1H), 10.82 (s, 1H), 12.29 (s, 1H).
Step B:
To a solution 4-hydroxynaphthoic acid in anhydrous methanol at 0xc2x0 C. was added thionyl chloride (1.5 eq). After stirring the solution at room temperature for 16 hours, the solvent was evaporated. The residue was taken up in ethyl acetate and washed with saturated aqueous sodium bicarbonate, water, brine, and dried over MgSO4 to give methyl 4-hydroxynaphthoate.
1H NMR (DMSO-D6): xcex4 3.87 (s, 3H), 6.92 (d, 1H), 7.53 (t, 1H), 7.65 (t, 1H), 8.13 (d, 1H), 8.26 (d, 1H), 8.93 (d, 1H), 11.16 (s, 1H).
Step C:
The title compound was prepared from methyl 4-hydroxynaphthoate according to the procedure for the synthesis of precursor hydrazides Axe2x80x94(Cxe2x95x90O)xe2x80x94NHNH2.
1H NMR (DMSO-D6): xcex4 6.60 (d, 1H), 7.28 (m, 3H), 7.95 (d, 1H), 8.07 (d, 1H), 9.25 (brd s, 1H).
Moreover, by use of the above methodology, the following hydrazides useful as intermediates in preparing the compounds of the invention may be prepared: 
The ether-linked aldehydes may be prepared by O-alkylation of the corresponding phenolic compounds using various electrophilic alkylating agents that introduce the xe2x80x94(K)mxe2x80x94D moiety as defined above in a reaction generally known as Williamson ether synthesis (H. Feuer, J. Hooz in The Chemistry of the Ether Linkage, S. Patai Ed., Wiley, New York 1967, p. 446-460). 
wherein Lx is a leaving group such as xe2x80x94Cl, xe2x80x94Br, xe2x80x94I, xe2x80x94OSO2CH3, xe2x80x94OSO2p-tolyl or xe2x80x94OSO2CF3; and
R3a, R3b, R4a, R4b, a, b, c, d, f, p, q, D, M, R14 and R15 are as defined for formula I.
According to Scheme II an ether-substituted aryl-aldehyde can be prepared by stirring hydroxybenzaldehydes or hydroxynaphthaldehydes in an organic solvent such as acetone, methylethyl ketone, dimethylformamide, dioxane, tetrahydrofuran, toluene, ethylene glycol dimethyl ether, sulfolane, diethylether, water or a compatible mixture of two or more of the above solvents with an equimolar amount of an alkyl halide or an aryl-lower alkyl halide and in the presence of 1 to 15 equivalents (preferably 1 to 5 equivalents) of a base such as sodium hydride, potassium hydride, sodium or potassium methoxide, ethoxide or tert-butoxide, sodium, potassium or cesium carbonate, potassium or cesium fluoride, sodium or potassium hydroxide or organic bases such as diisopropylethylamine, 2,4,6-collidine or benzyldimethyl-ammonium methoxide or hydroxide. The reaction can be performed at 0xc2x0 C. to 150xc2x0 C., preferably at 20xc2x0 C. to 100xc2x0 C. and preferably in an inert atmosphere of N2 or Ar. When the reaction is complete the mixture is filtered, concentrated in vacuo and the resulting product optionally purified by column chromatography on silica gel using ethyl acetate/hexane as eluent. The compound can also (when appropriate) be purified by recrystallization from a suitable solvent such as ethyl alcohol, ethyl acetate, isopropyl alcohol, water, hexane, toluene or their compatible mixture. Specific examples illustrating the preparation of ether-substituted aryl-aldehydes are provided below.
A mixture of 4-hydroxynaphthaldehyde (1 g, 5.8 mmol), 2-bromomethyl tetrahydropyran (1 g, 5.8 mmol) and powdered K2CO3 (1.2 g, 8.7 mmol) in dimethyl formamide was stirred at 60xc2x0 C. overnight. The mixture was taken up in water and ethyl acetate. The organic layer was separated and washed with water, brine, dried over MgSO4, filtered, and concentrated. The product was purified by silica gel column chromatography using ethyl acetate/hexane. 
1H NMR (DMSO-d6): xcex4 1.48 (m, 4H), 1.74 (d, 1H), 1.84 (m, 1H), 3.44 (m, 1H), 3.78 (m, 1H), 3.92 (d, 1H), 4.23 (m, 2H), 7.17 (d, 1H), 7.64 (t, 1H), 7.74 (t, 1H), 8.11 (d, 1H), 8.27 (d, 1H), 9.22 (d, 1H), 10.17 (s, 1H).
A mixture of 4-hydroxynaphthaldehyde (1 g, 5.8 mmol), 3,5-bis-trifluoromethylbenzylbromide (1.8 g, 5.8 mmol), and powdered K2CO3 (1.2 g, 8.7 mmol) was stirred in acetone (40 mL) overnight. The mixture was poured onto 200 mL of ice-chips and stirred until the ice melted. The yellow precipitate, 4-((3,5-bis-trifluoromethyl)benzyloxy)-1-naphthaldehyde, was collected and dried. 
1H NMR (DMSO-d6): xcex4 5.58 (s, 2H), 7.07 (d, 1H), 7.22 (d, 1H), 7.63 (t, 1H), 7.69 (t, 1H), 7.79 (d, 1H), 7.86 (d, 1H), 7.99 (s, 1H), 8.14 (s, 1H), 8.30 (s, 3H), 8.94 (s, 1H), 8.97 (d, 1H), 11.0 (broad s, 1H), 11.69 (s, 1H); MS (ESI) m/z 675.2 (M+H)+.
To a solution of 4-hydroxy-1-naphthaldehyde (8.6 g, 50 mmoles) and potassium carbonate (13.8 g, 100 mmoles) in N,N-dimethylformamide (DMF)(40 mL) was added 1-bromo-2-chloroethane (7.4 g, 50 mmoles). The mixture was heated at 60xc2x0 C. overnight. The solution was diluted with ethyl acetate (500 mL), extracted with water and brine. The organic layer was dried over magnesium sulfate and the solvent was evaporated to obtain 12.1 g product (52% yield). 
MS (Cl): 403, 405, 407. 1H NMR (CDCl3): xcex4 10.2 (s, 1H), 9.3 (d, 1H), 8.35 (d, 1H), 7.85 (d, 1H), 7.65 (m, 1H), 7.5 (m, 1H), 7.1 (d, 1H), 4.35 (t, 2H), 4.15 (t, 2H).
The products were used as such in further transformations.
By application of the above methodology the following substituted aldehyde intermediates were synthesized: 

To a solution of vanillin (1.0 g, 6.57 mmol) in acetone (30 mL) was added potassium carbonate (4.50 g, 32.8 mmol) and allyl bromide (0.62 mL, 7.3 mmol). The mixture was heated under reflux for 6 h. TLC showed appearance of a new spot. Potassium salts were removed by filtration and the filtrate was concentrated to a syrup. A small sample was purified using prep TLC using hexane/ethyl acetate 7:3 as developing solvent. 1H NMR (CDCl3) xcex4=3.94 (s, 3H), 4.67-4.83 (m, 2H), 5.30-5.55 (m, 2H), 6.01-6.21 (m, 1H), 6.98 (d, J=9 Hz, 1H), 7.40-7.56 (m, 2H), 9.85 (s, 1H); MS (APCI): 193.6.
The crude syrup was heated neat in an oil bath at 200xc2x0 C. for 6 h. The crude material was dissolved in chloroform and filtered through a pack of silica gel. The crude product (yield 72%) was used as is in the next step for O-alkylation. A small portion was purified using prep-TLC to give a pure sample of 3-allyl-4-hydroxy-5-methoxy-benzaldehyde. 1H NMR (CDCl3) xcex4=3.46 (d, J=6 Hz, 2H), 3.96 (s, 3H), 5.02-5.22 (m, 2H), 5.94-6.11 (m, 1H), 6.30 (s, 1H), 7.45 (s, 2H), 9.80 (s, 1H); MS (APCI): 193.3.
The crude 3-allyl-4-hydroxy-5-methoxy-benzaldehyde was taken up in acetone and treated with 4-isopropylbenzyl chloride in the presence of potassium carbonate to give the desired product. 
1H NMR (CDCl3) xcex4=1.26 (d, J=7 Hz, 6H), 2.92 (m, 1H), 3.38 (d, J=7 Hz, 2H), 3.95 (s, 3H), 4.98-5.12 (m, 4H), 5.93-5.75 (m, 1H), 7.20-7.43 (m, 6H), 9.87 (s, 1H).

In the above formulae B, D, R8 and R9 have the same meanings as defined for formula I.
Step A:
To a solution of aniline (or an aniline derivative) (1 eq.) in THF was added dropwise chloroacetyl chloride (1.2 eq.). After stirring at room temperature overnight, 100 mL water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed twice with dilute hydrochloric acid, twice with water, dried over Mg SO4 and then concentrated to give pure product.
Step B:
To a solution of chloroacetanilide (or a derivative thereof) (1.2 eq.) and 2-methoxy-4-hydroxy benzaldehyde (or another aromatic aldehyde substituted with a hydroxy group) (1 eq.) in DMSO was added potassium carbonate (1.5 eq.). After stirring overnight at room temperature, 100 ml water was added. The mixture was extracted with ethyl acetate, the organic extracts were washed twice with a satd. sodium bicarbonate solution, twice with water, and dried over MgSO4. After concentration in vacuo, the product was obtained.
The following two aldehydes were prepared as examples of compounds that can be prepared using this methodology:
N-(4-Chlorophenyl)-2-(4-formyl-3-methoxyphenoxy)acetamide: 
1H NMR (CDCl3): xcex4 4.28 (s, 3H), 5.01 (s, 2H), 6.90 (d, J=2.2 Hz, 1H), 6.97 (dd, J=8.6, 2.1 Hz, 1H), 7.67 (d, J=8.9 Hz, 2H), 7.89 (d, J=8.8 Hz, 2H), 8.20 (d, J=8.6 Hz, 1H), 8.51 (s, 1H), 10.66 (s, 1H); MS (APCI): 319.9.
N-(4-Isopropylphenyl)-2-(4-formyl-3-methoxyphenoxy)acetamide: 
1H NMR (DMSO-D6): xcex4 2.07 (d, J=6.9 Hz, 6H), 2.70 (m, J=6.9 Hz, 1H), 3.77 (s, 3H), 4.68 (s, 2H), 6.56 (dd, J=8.7, 2.1 Hz, 1H), 6.66 (d, J=2.1 Hz, 1H), 7.06 (d, J=8.5 Hz, 2H), 7.39 (d, J=8.50 Hz, 2H), 7.55 (d, J=8.7 Hz, 1H), 9.93 (s, 1H), 10.05 (s, 1H); MS (APCI): 328.
This type of aldehydes can be coupled to hydrazides using the methodology as described in step D to give a compound of formula IXa. Alternatively these compounds can undergo rearrangement by treatment with base as described below (step C), followed by coupling to a hydrazide (step D) to give a compound of formula IXb.
Step C:
The mixture of aldehyde (1 eq.) and potassium carbonate (1.5 eq.) in acetonitrile was refluxed. The reaction was monitored by TLC (hexane:ethyl acetate=2:1). When TLC showed almost complete conversion (about 48 h), 100 ml water was added. The mixture was extracted with ethyl acetate, the organic extracts were dried over MgSO4, and concentrated to give the desired product which can be further purified by column chromatography, or used directly for the next step.
The following two aldehydes were prepared as examples of compounds that can be prepared using this methodology:
4-(4-Chlorophenylamino)-2-methoxybenzaldehyde:
Prepared from N-(4-chlorophenyl)-2-(4-formyl-3-methoxyphenoxy)acetamide using the procedure described in step C above. 
1H NMR (CDCl3): xcex4 3.84 (s, 3H), 6.14 (s, 1H), 6.45 (d, J=2.0 Hz, 1H), 6.54 (dd, J=8.4, 1.8 Hz, 1H), 7.14 (d, J=8.7 Hz, 2H), 7.33 (d, J=8.7 Hz, 2H), 7.74 (d, J=8.5 Hz, 1H), 10.22 (s, 1H); MS (APCI): 261.9.
4-(4-Isopropylphenylamino)-2-methoxybenzaldehyde:
Prepared from N-(4-isopropylphenyl)-2-(4-formyl-3-methoxyphenoxy)acetamide using the procedure described in step C above. 
1H NMR (CDCl3) xcex4 1.26 (d, J=6.9 Hz, 6H), 2.88 (m, J=6.9 Hz, 1H), 3.84 (s, 3H), 6.50 (d, J=1.9 Hz, 1H), 6.55 (dd, J=8.6, 1.8 Hz, 1H), 6.96 (s, 1H), 7.15 (d, 2H, J=8.5 Hz, 2H), 7.22 (d, J=8.5 Hz, 2H), 7.69 (d, J=8.5 Hz, 1H), 10.18 (s, 1H); MS (APCI): 269.
Step D:
The resulting carbonyl compounds are treated with the corresponding acylhydrazide in a solvent. The solvent may be one of the following:ethyl alcohol, methyl alcohol, isopropyl alcohol, tert-butyl alcohol, dioxane, tetrahydrofuran, toluene, chlorobenzene, anisole, benzene, chloroform, dichloromethane, DMSO, acetic acid, water or a compatible mixture of two or more of the above solvents. A catalyst such as acetic acid can be added. A dehydrating reagent such as triethylorthoformate can also be added to the reaction mixture. The reaction is performed by stirring the reaction mixture preferably under an inert atmosphere of N2 or Ar at temperatures between 0xc2x0 C. to 140xc2x0 C., preferably between 10xc2x0 C. to 80xc2x0 C. In many cases the product simply crystallizes out when the reaction is completed and is isolated by suction filtration. It can be further recrystallized if necessary from a solvent such as the above described reaction solvents. The product can also be isolated by concentration of the reaction mixture in vacuo, followed by column chromatography on silica gel using a solvent system such as chloroform/methanol or dichloromethane/methanol or chloroform/ethyl acetate to give a compound of formula IXb.